LOCUS       sequence-217841-       10535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for FLPe recombinase-mediated cassette exchange in 
            master cell lines created with plasmid #112666. This vector allows 
            inducible expression of your protein of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10535)
  AUTHORS   Ordovas L, Boon R, Pistoni M, Chen Y, Wolfs E, Guo W, Sambathkumar 
            R, Bobis-Wozowicz S, Helsen N, Vanhove J, Berckmans P, Cai Q, 
            Vanuytsel K, Eggermont K, Vanslembrouck V, Schmidt BZ, Raitano S, 
            Van Den Bosch L, Nahmias Y, Cathomen T, Struys T, Verfaillie CM
  TITLE     Efficient Recombinase-Mediated Cassette Exchange in hPSCs to Study 
            the Hepatocyte Lineage Reveals AAVS1 Locus-Mediated Transgene 
            Inhibition.
  JOURNAL   Stem Cell Reports. 2015 Nov 10;5(5):918-931. doi: 
            10.1016/j.stemcr.2015.09.004. Epub 2015 Oct 8.
  PUBMED    26455413
REFERENCE   2  (bases 1 to 10535)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.stemcr.2015.09.004"; journalName: "Stem Cell Reports"; 
            date: "2015-11-10- 10"; volume: "5"; issue: "5"; pages: "918-931"
COMMENT     Sequence Label: pZ P TetON-3XFLAG-tdT CAGG-m2rtTA v2
FEATURES             Location/Qualifiers
     source          1..10535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    659..1462
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    1469..1502
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    1506..1531
                     /label=SA
                     /note="splice acceptor site"
     CDS             1552..1605
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1615..2214
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1615..1634)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2111..2131
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2368..2479
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2734..2781
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2806..2825
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2845..4275)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     regulatory      4272..4281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4365..4374
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4382..4406)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4403..4441)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    4453..4471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4489..4507
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4524..4542
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4560..4578
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4596..4614
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4631..4649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     enhancer        4710..5093
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5098..5375
                     /label=chicken beta-actin promoter
     intron          5376..6392
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     6316..6338
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     6400..6419
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(6447..7064,7065..7070,7071..7106,7107..7109,7110..7145,
                     7146..7148,7149..7184,7185..7193)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    7207..7431
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    7438..7471
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    7481..8317
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(8347..8367)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8347..8365)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8386..8402)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8386..8402)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8399..8421)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8410..8426
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8434..8440,8441..8458,8459..8464))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8479..8500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8617..8634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8788..9376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8868..8887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9547..10338,10339..10407))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10170..10189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10408..10512)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacctg
      661 ctttctctga ccagcattct ctcccctggg cctgtgccgc tttctgtctg cagcttgtgg
      721 cctgggtcac ctctacggct ggcccagatc cttccctgcc gcctccttca ggttccgtct
      781 tcctccactc cctcttcccc ttgctctctg ctgtgttgct gcccaaggat gctctttccg
      841 gagcacttcc ttctcggcgc tgcaccacgt gatgtcctct gagcggatcc tccccgtgtc
      901 tgggtcctct ccgggcatct ctcctccctc acccaacccc atgccgtctt cactcgctgg
      961 gttccctttt ccttctcctt ctggggcctg tgccatctct cgtttcttag gatggccttc
     1021 tccgacggat gtctcccttg cgtcccgcct ccccttcttg taggcctgca tcatcaccgt
     1081 ttttctggac aaccccaaag taccccgtct ccctggcttt agccacctct ccatcctctt
     1141 gctttctttg cctggacacc ccgttctcct gtggattcgg gtcacctctc actcctttca
     1201 tttgggcagc tcccctaccc cccttacctc tctagtctgt gctagctctt ccagccccct
     1261 gtcatggcat cttccagggg tccgagagct cagctagtct tcttcctcca acccgggccc
     1321 ctatgtccac ttcaggacag catgtttgct gcctccaggg atcctgtgtc cccgagctgg
     1381 gaccacctta tattcccagg gccggttaat gtggctctgg ttctgggtac ttttatctgt
     1441 cccctccacc ccacagtggg gcaagcttga agttcctatt cttcaaatag tataggaact
     1501 tcggcctgac ctcttctctt cctcccacag ggcctcagat ctggcagcgg agagggcaga
     1561 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggct cgaaatgacc
     1621 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc
     1681 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac
     1741 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     1801 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     1861 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     1921 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     1981 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     2041 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     2101 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     2161 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     2221 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc cgaccgaagc
     2281 cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgac tctagagggc
     2341 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2401 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2461 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2521 tggggcagga cagcaagggg gaggattggg aagacaatag cactcgagct atggcagggc
     2581 ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg
     2641 gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag
     2701 agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt
     2761 tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg
     2821 tttcagttag cctcccccac gcgtttactt gtacagctcg tccatgccgt acaggaacag
     2881 gtggtggcgg ccctcggagc gctcgtactg ttccacgatg gtgtagtcct cgttgtggga
     2941 ggtgatgtcc agcttggtgt ccacgtagta gtagccgggc agttgcacgg gcttcttggc
     3001 catgtagatg gtcttgaact ccaccaggta gtggccgccg tccttcagct tcagggcctg
     3061 gtggatctcg cccttcagca cgccgtcgcg ggggtacagg cgctcggtgg aggcctccca
     3121 gcccatggtc ttcttctgca ttacggggcc gtcggggggg aagttggtgc cgcgcatctt
     3181 caccttgtag atcagcgtgc cgtcctgcag ggaggagtcc tgggtcacgg tcaccagacc
     3241 gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg acagcttctt
     3301 gtaatcgggg atgtcggcgg ggtgcttcac gtacgccttg gagccgtaca tgaactgggg
     3361 ggacaggatg tcccaggcga agggcagggg gccgcccttg gtcaccttca gcttggcggt
     3421 ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact cgtggccgtt
     3481 catggagccc tccatgcgca ccttgaagcg catgaactct ttgatgacgg ccatgttgtt
     3541 gtcctcggag gaggcggtgc cggagctgcc gctgccggtg ctgccggtgc catgccccag
     3601 gaacaggtgg tggcggccct cggagcgctc gtactgttcc acgatggtgt agtcctcgtt
     3661 gtgggaggtg atgtccagct tggtgtccac gtagtagtag ccgggcagtt gcacgggctt
     3721 cttggccatg tagatggtct tgaactccac caggtagtgg ccgccgtcct tcagcttcag
     3781 ggcctggtgg atctcgccct tcagcacgcc gtcgcggggg tacaggcgct cggtggaggc
     3841 ctcccagccc atggtcttct tctgcattac ggggccgtcg ggggggaagt tggtgccgcg
     3901 catcttcacc ttgtagatca gcgtgccgtc ctgcagggag gagtcctggg tcacggtcac
     3961 cagaccgccg tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg ggaaggacag
     4021 cttcttgtaa tcggggatgt cggcggggtg cttcacgtac gccttggagc cgtacatgaa
     4081 ctggggggac aggatgtccc aggcgaaggg cagggggccg cccttggtca ccttcagctt
     4141 ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg
     4201 gccgttcatg gagccctcca tgcgcacctt gaagcgcatg aactctttga tgacctcctc
     4261 gcccttgctc accatggtgg catatggata tctccaccgg tcgtcatcgt catccttgta
     4321 atcaatatca tgatctttat aatcaccgtc atggtctttg tagtccatgg tggccttaag
     4381 cgatctgacg gttcactaaa cgagctctgc ttatataggc ctcccaccgt acacgcctac
     4441 ctcgacatac gttctctatc actgataggg agtaaactcg acatacgttc tctatcactg
     4501 atagggataa actcgacata cgttctctat cactgatagg gagtaaactc gacatacgtt
     4561 ctctatcact gatagggagt aaactcgaca tacgttctct atcactgata gggagtaaac
     4621 tcgacatcgt tctctatcac tgatagggag taaactcgtc tagaacctcg gaccttcgaa
     4681 aaaagtttcg agtgtacagc atcatcgtcg acattgatta ttgactagct agttattaat
     4741 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4801 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4861 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4921 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4981 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     5041 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgatggtcg
     5101 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     5161 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc
     5221 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
     5281 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     5341 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
     5401 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     5461 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
     5521 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
     5581 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
     5641 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
     5701 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     5761 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     5821 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     5881 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     5941 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     6001 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     6061 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     6121 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     6181 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     6241 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     6301 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     6361 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     6421 tgtctcatca ttttggcaaa gaattcatgt ctagactgga caagagcaaa gtcataaacg
     6481 gcgctctgga attactcaat ggagtcggta tcgaaggcct gacgacaagg aaactcgctc
     6541 aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag cgggccctgc
     6601 tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc cccctggaag
     6661 gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc attccgctgt gctctcctct
     6721 cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa
     6781 ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag aacgcactgt
     6841 acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa caggagcatc
     6901 aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca cttctgagac
     6961 aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc ggcctggaac
     7021 taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg gccgacgccc
     7081 ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt gaccttgata
     7141 tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg taaactagtg
     7201 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     7261 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     7321 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     7381 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gttcgaagaa
     7441 gttcctattc tctagaaagt ataggaactt cgcggccgct actagggaca ggattggtga
     7501 cagaaaagcc ccatccttag gcctcctcct tcctagtctc ctgatattgg gtctaacccc
     7561 cacctcctgt taggcagatt ccttatctgg tgacacaccc ccatttcctg gagccatctc
     7621 tctccttgcc agaacctcta aggtttgctt acgatggagc cagagaggat cctgggaggg
     7681 agagcttggc agggggtggg agggaagggg gggatgcgtg acctgcccgg ttctcagtgg
     7741 ccaccctgcg ctaccctctc ccagaacctg agctgctctg acgcggctgt ctggtgcgtt
     7801 tcactgatcc tggtgctgca gcttccttac acttcccaag aggagaagca gtttggaaaa
     7861 acaaaatcag aataagttgg tcctgagttc taactttggc tcttcacctt tctagtcccc
     7921 aatttatatt gttcctccgt gcgtcagttt tacctgtgag ataaggccag tagccagccc
     7981 cgtcctggca gggctgtggt gaggaggggg gtgtccgtgt ggaaaactcc ctttgtgaga
     8041 atggtgcgtc ctaggtgttc accaggtcgt ggccgcctct actccctttc tctttctcca
     8101 tccttctttc cttaaagagt ccccagtgct atctgggaca tattcctccg cccagagcag
     8161 ggtcccgctt ccctaaggcc ctgctctggg cttctgggtt tgagtccttg gcaagcccag
     8221 gagaggcgct caggcttccc tgtccccctt cctcgtccac catctcatgc ccctggctct
     8281 cctgcccctt ccctacaggg gttcctggct ctgctctcca ccgcggtgga gctccagctt
     8341 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     8401 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     8461 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     8521 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     8581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     8641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     8701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     8761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     8821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     8881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     8941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     9001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     9061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     9121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     9181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     9241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     9301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     9361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     9421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     9481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     9541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     9601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     9661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     9721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     9781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     9841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     9901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     9961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
    10021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
    10081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
    10141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    10201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    10261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    10321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    10381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    10441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
    10501 aggggttccg cgcacatttc cccgaaaagt gccac
//
